Evidence that tetrahydrofolate does not bind to serine hydroxymethyltransferase with positive homotropic cooperativity.
Previous experiments suggesting that tetrahydrofolate binds to serine hydroxymethyltransferase with positive homotropic cooperativity have been reinvestigated. Our results show that the sigmoid-shaped tetrahydrofolate saturation curve, previously obtained by several other investigators, is due to the instability of tetrahydrofolate in the assay solution. Using a different assay method, we have shown that tetrahydrofolate gives a hyperbolic saturation curve with serine hydroxymethyltransferase. We could find no evidence, as suggested by other investigators, that heating the enzyme during purification destroyed its allosteric properties or that NADH binds to the enzyme as an allosteric effector. Evidence is presented that the loss of tetrahydrofolate during the assay period is due to oxidation by dissolved molecular oxygen.
D000818:Animals; D004355:Drug Stability; D012696:Glycine Hydroxymethyltransferase / Q000378:metabolism*; D000882:Haplorhini; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D009243:NAD / Q000494:pharmacology; D010084:Oxidation-Reduction; D011485:Protein Binding / Q000187:drug effects; D011817:Rabbits; D013763:Tetrahydrofolates / Q000378:metabolism*; D014166:Transferases / Q000378:metabolism*
D013763:Tetrahydrofolates; D009243:NAD; C030371:5,6,7,8-tetrahydrofolic acid; D014166:Transferases; D012696:Glycine Hydroxymethyltransferase